Polymerisation of gluten proteins in developing wheat grain as affected by desiccation

The breadmaking quality of wheat is affected by the composition of gluten proteins and the polymerisation of subunits that are synthesised and accumulated in developing wheat grain. The biological mechanisms and time course of these events during grain development are documented, but not widely confirmed. Therefore, the aim of this study was to monitor the accumulation of gluten protein subunits and the size distribution of protein aggregates during grain development. The effect of desiccation on the polymerisation of gluten proteins and the functional properties of gluten were also studied. The results showed that the size of glutenin polymers remained consistently low until yellow ripeness (YR), while it increased during grain desiccation after YR. Hence, this polymerisation process was presumed to be initiated by desiccation. A similar polymerisation event was also observed when premature grains were dried artificially. The composition of gluten proteins, the ratios of glutenin to gliadin and high molecular weight-glutenin subunits to low molecular weight-glutenin subunits, in premature grain after artificial desiccation showed close association with the size of glutenin polymers in artificially dried grain. Functional properties of gluten in these samples were also associated with polymer size after artificial desiccation.     

Impact of ethanol,succinic acid,and the combination thereof at levels produced during sponge fermentation on hard wheat,soft wheat,and durum wheat farinograph rheology

The sponge and dough mixing process is one of the most common in the world, yet the mechanistic understanding of this process has yet to be sufficiently explored. In this study, aqueous solutions of ethanol, succinic acid, and their combination were prepared at concentrations intended to replicate fermentation times of 3, 4 and 6 h. These solutions were added to a farinograph mixer to make dough using hard wheat, soft wheat, and durum wheat flour. The results indicate that these yeast metabolites (ethanol, succinic acid) impact the mixing resistance, peak mixing value, and dough mixing stability in each of the flour types, likely primarily affected by the ratios of gliadin to glutenin and LMW glutenin in each flour type. Results suggest a stabilizing non-covalent interaction imparted by gliadin at peak mixing time, a stabilizing effect of HMW glutenin during break down, and synergistic effects of ethanol and succinic acid that leads to a faster rate of breakdown in later stages of mixing. It also suggests an increase in mixing resistance when acidulants are added to durum wheat dough. Taken together, this study adds new insights on the sponge and dough mixing process in a way that has not previously been conducted.     

Extractability and chromatographic separation of rye (Secale cereale L.) flour proteins

A size exclusion – high performance liquid chromatography (SE-HPLC) method originally developed for separating wheat, barley or rice proteins was applied to study the extractability and molecular weight (MW) distribution of rye flour proteins. These were extracted with 50 mmol/l sodium phosphate buffer (pH 6.8) containing 2.0% (w/v) sodium dodecyl sulfate (SDS) and, optionally, 1.0% (w/v) dithiothreitol (DTT). About 95% of the proteins were extracted in buffer containing 2.0% SDS. Addition of 1.0% DTT to such buffer increased the protein extractability to 100%, indicating that rye flour contains some proteins cross-linked by disulfide (SS) bonds. The SE-HPLC profiles revealed that rye flour contains SS-linked HMW-secalins and 75 k γ-secalins which elute in specific peaks. Upon reduction, these SS-linked protein aggregates dissociate and some entrapped albumins, globulins and/or ω-secalins are released. Rye flour albumins and globulins elute over the entire SE-HPLC profile. In contrast, the monomeric ω-secalins and 40 k γ-secalins are detected in specific well resolved SE-HPLC peaks. The applied fast and reproducible method can be used to characterise and quantify rye flour proteins and to determine changes as a result of processing.     

Study of the mechanism of improvement due to waxy wheat flour addition on the quality of frozen dough bread

Waxy wheat flour (WWF) was substituted for 10% regular wheat flour (RWF) in frozen doughs and the physicochemical properties of starch and protein isolated from the frozen doughs stored for different time intervals (0, 1, 2, 4 and 8 weeks) were determined to establish the underlying reasons leading to the effects observed in WWF addition on frozen dough quality. Using Nuclear Magnetic Resonance (NMR), Differential Scanning Calorimeter (DSC) and X-ray Diffraction (XRD) among others, the gluten content, water molecular state, glutenin macropolymer content, damaged starch content, starch swelling power, gelatinization properties, starch crystallinity and bread specific volume were measured. Compared to RWF dough at the same frozen storage condition, 10% WWF addition decreased dry gluten and glutenin macropolymer contents and T₂₃ proton density of frozen dough, but increased the wet gluten content, T₂₁ and T₂₂ proton density. 10% WWF addition also decreased damaged starch content, but increased starch swelling power, gelatinization temperature and enthalpy, crystallinity of starch and bread specific volume of frozen dough. Results in the present study showed that the improvement observed due to WWF addition in frozen dough bread quality might be attributed to its inhibition of redistribution of water molecules bound to proteins, increase in damaged starch content and decrease in starch swelling power.     

GlutoPeak parameters of whole wheat flours for gluten quality evaluation in soft wheat breeding programs

In soft wheat breeding programs, the gluten strength of flours from specific genotypes is determined by various chemical and rheological tests. Based on such tests, the experimental wheat lines with very weak flour gluten are typically selected for the production of soft-dough biscuits, while the lines with medium gluten strength and extensibility are reserved for hard-dough biscuits. Often, the genotypes having high gluten strength are removed from such breeding programs. In the present study, the usability of the GlutoPeak tester on whole wheat flour samples was investigated for assessing the gluten strength of soft wheat breeding materials. In the study, 25 soft wheat genotypes, grown in seven locations for three years, were categorized by commonly used gluten-quality-related parameters. Based on the results of the study GlutoPeak whole wheat flour PMT values ranging from 30.0 to 50.0 s and AM values from 15.0 to 20.0 GPU were found to be suitable for soft-dough biscuit products, whereas the values between 40.0 and 60.0 s and 20.0 and 23.0 GPU were appropriate for hard-dough biscuit products. The genotypes exhibiting AM values > 24.0 GPU and PMT values > 60.0 s were judged to have too-strong gluten, and thus eliminated from the breeding program. The gluten aggregation energy (AGGEN), and the torque after the maximum torque (PM) values were only useful and applicable to flours for soft-dough products. The maximum torque (BEM) values were not effective in discriminating against the genotypes. The results of this study demonstrated that the GlutoPeak whole wheat PMT and AM parameters can be recommended as quick and accurate parameters especially for early generation screening with small-scale tests in soft wheat improvement programs.     

The impact of Septoria tritici blotch in bread making quality among argentinean wheat cultivars

Septoria tritici blotch (STB), caused by Zymoseptoria tritici is a relevant foliar wheat disease worldwide. Several reports show the importance of STB on grain yield, their components and grain protein while little is known about its effect on the rheological properties of the wheat flour. The scarce literature found, only mentions the effect of the complex of foliar diseases on wheat quality, without individualizing the effect of the different pathogens separately. This study analyze the influence of increasing doses of inoculum of Zymoseptoria tritici, on the bread making quality of ten Argentinean wheat cultivars and its possible variation according to their quality group. The increase of inoculum concentration augmented the area under disease progress curve, decreased green flag leaf area duration and green leaf area duration. Cultivars K. Flecha and B.75 Aniversario had the lowest green flag leaf area duration causing higher reduction in grain filling period and higher reductions in P, indicating a lower gliadin/glutenin ratio. STB decreased P/L and E while L, W, D, SV and bread volume increased. Cultivars differed in rheological parameters according to their quality group. Gluten/protein relationship was significant in quality group 1 and non-significant in cultivars belonging to quality group 2 and 3.     

Effects of wheat tempering and stone rotational speed on particle size,dough rheology and bread characteristics for a stone-milled weak flour

The poor technological performance of weak wheat flours means that they are usually considered difficult to be transformed into satisfactory bread. During milling, there are several settings that can affect flour characteristics. In this study, we tested two operative parameters that have the potential to affect flour quality – stone rotational speed and wheat tempering. Tempering moistures were set at 11%, 13%, 15%, and 17%, while stone rotational speeds were set at 173, 260, and 346 rpm. Both factors were found to affect operative milling parameters, notably flour yield, process productivity and specific energy consumption. Grain moisture had a significant effect on both dough rheology and bread characteristics (dough stability, tenacity, and extensibility). Dough stability was maximum at 13% moisture. Dough tenacity decreased as moisture increased, while extensibility increased as moisture increased. Bread specific volume and crumb specific volume were improved at 13% and 15% moisture. In conclusion, wheat tempering can be used to improve the potential of a weak flour and bread characteristics, while stone rotational speed affects operative parameters and white flour yield.     

Transgenic sorghum with suppressed synthesis of kafirin subclasses: Effects on flour and dough rheological characteristics

Arising from work showing that conventionally bred high protein digestibility sorghum types have improved flour and dough functionality, the flour and dough properties of transgenic biofortified sorghum lines with increased protein digestibility and high lysine content (TG-HD) resulting from suppressed synthesis of several kafirin subclasses, especially the cysteine-rich γ-kafirin, were studied. TG-HD sorghums had higher flour water solubility at 30 °C (p < 0.05) and much higher paste viscosity (41% higher) than their null controls (NC). TG-HD doughs were twice as strong as their NC and dynamic rheological analysis indicated that the TG doughs were somewhat more elastic up to 90 °C. CLSM of doughs and pastes indicated that TG-HD had a less compact endosperm protein matrix surround the starch compared to their NC. The improved flour and dough functional properties of the TG-HD sorghums seem to be caused by reduced endosperm compactness resulting from suppression of synthesis of several kafirin subclasses which modifies protein body and protein matrix structure, and to improved protein-starch interaction through hydrogen bonding specifically caused by reduction in the level of the hydrophobic γ-kafirin. The improved flour functionality of these transgenic biofortified sorghums can increase their commercial utility by complementing their improved nutritional quality.     

Combined mutations in five wheat STARCH BRANCHING ENZYME II genes improve resistant starch but affect grain yield and bread-making quality

Increases in the proportion of amylose in the starch of wheat grains result in higher levels of resistant starch, a fermentable dietary fiber associated with human health benefits. The objective of this study was to assess the effect of combined mutations in five STARCH BRANCHING ENZYME II (SBEII) genes on starch composition, grain yield and bread-making quality in two hexaploid wheat varieties. Significantly higher amylose (∼60%) and resistant starch content (10-fold) was detected in the SBEII mutants than in the wild-type controls. Mutant lines showed a significant decrease in total starch (6%), kernel weight (3%) and total grain yield (6%). Effects of the mutations in bread-making quality included increases in grain hardness, starch damage, water absorption and flour protein content; and reductions in flour extraction, farinograph development and stability times, starch viscosity, and loaf volume. Several traits showed significant interactions between genotypes, varieties, and environments, suggesting that some of the negative impacts of the combined SBEII mutations can be ameliorated by adequate selection of genetic background and growing location. The deployment of wheat varieties with increased resistant starch will likely require economic incentives to compensate growers and millers for the significant reductions detected in grain and flour yields.     

Predicting water absorption of wheat flour using high shear-based GlutoPeak test

Selection for water absorption, a fundamental wheat quality parameter, has been a challenge in wheat breeding programs due to limited wheat materials available for milling and consequent time-consuming farinograph test. Hence, a high shear-based method, which requires 8 g of flour and less than 10 min per test, was proposed to predict flour water absorption using the Brabender GlutoPeak instrument. Highly significant positive linear relationship (r² = 0.97) was found between GlutoPeak maximum torque and farinograph water absorption for 83 flour samples prepared with Bühler test mill from wheat lines under evaluation in the Canadian wheat variety registration trials. Similar strong correlation (r² = 0.96) was obtained from flours (n = 63) prepared with Quadrumat Junior laboratory mill using small amount of wheat. Flour prepared either with Bühler test mill or Quadrumat Junior mill can be used for predicting water absorption effectively. GlutoPeak maximum torque was found to be independent of dough strength (r² = 0.02) as measured by extensigraph. GlutoPeak test can be a powerful tool for rapid and reliable prediction of water absorption of wheat flour.     

Gluten proteome comparison among durum wheat genotypes with different release date

In order to deepen insight into durum wheat prolamin composition in relation to both end use quality and gluten related disorders, a phenotyping of four genotypes of different release date and technological performance was carried out. A proteomic approach integrated with the evaluation of protein aggregation level, amino acid composition and reactivity to G12 monoclonal antibody, was adopted. The degree of polymerization, estimated as unextractable polymeric proteins (UPP), was positively influenced by Cys-rich proteins (Glu-B3 LMW-GS), with Saragolla showing up to 40% higher values than other genotypes. The proteomic assessment allowed to identify proteins involved in gluten related disorders. In particular, ω5-gliadin involved in wheat allergy (WA), resulted markedly over-expressed in the old landrace Dauno III, up to four-fold than in modern Saragolla. A marked influence of genotype on the expression level of gliadins containing toxic epitopes (TECP) was observed with the more recent genotypes showing lower values (−53%). Also, reactivity to G12 moAb resulted higher in the two old genotypes consistently to their higher celiac disease (CD) toxicity evaluated by the proteomic approach. In conclusion a better gluten composition for processing seems to be associated to a lower expression level of CD toxic peptides and Tri a 19.     

Determination of volatile compounds in heat-treated straight-grade flours from normal and waxy wheats

Volatile compounds formed during heat-treatment of wheat flour influence the application of treated flour. In this study, normal and waxy hard wheat flours before and after dry-heat treatment were subjected to headspace analysis by solid-phase microextraction of volatiles followed gas chromatography–mass spectrometry (GC/MS). The untreated waxy wheat flour contained higher levels of odor-active compounds than normal wheat flour including aldehydes, alcohols, furans, and ketones. Lipid oxidation appears to play major role in producing such odor compounds. Heat treatments, depending on the severity, alter the profile of volatile compounds. Low temperature (100–110 °C) treatments effectively eliminated cereal odor (aldehyde) and did not introduce additional odors, providing a possible way to produce low-odor flours. Heat treatments at 120 °C and higher temperatures elevated the content of pyrazines, furans, and sulfur-containing compounds which together gave a roasty aroma to the flours. Considering organoleptic properties, treatments of flours at 140 °C was superior to 160 °C. The waxy wheat flour was more prone to produce odor-active compounds than normal wheat flour during the same heat treatment.     

Whole wheat bread: Effect of bran fractions on dough and end-product quality

Consumption of whole-wheat based products is encouraged due to their important nutritional elements that benefit human health. However, the use of whole-wheat flour is limited because of the poor processing and end-product quality. Bran was postulated as the major problem in whole wheat breadmaking. In this study, four major bran components including lipids, extractable phenolics (EP), hydrolysable phenolics (HP), and fiber were evaluated for their specific functionality in flour, dough and bread baking. The experiment was done by reconstitution approach using the 2⁴ factorial experimental layout. Fiber was identified as a main component to have highly significant (P < 0.05) and negative influence on most breadmaking characteristics. Although HP had positive effect on farinograph stability, it was identified as another main factor that negatively impacted the oven spring and bread loaf volume. Bran oil and EP seemed to be detrimental to most breadmaking characteristics. Overall, statistical analysis indicates that influence of the four bran components are highly complex. The bran components demonstrate multi-way interactions in regards to their influence on dough and bread-making characteristics. Particularly, Fiber appeared to have a high degree of interaction with other bran components and notably influenced the functionality of those components in whole wheat bread-making.     

Modification of physicochemical properties and in vitro digestibility of wheat flour through superheated steam processing

Native and moistened wheat flours (moisture contents were 13.5 and 27.0%, respectively) were treated with superheated steam (SS) at different temperatures (140 and 170 °C) and times (1, 2 and 4 min). Their physicochemical and digestive properties were analyzed. For native flour, SS treatment altered the starch molecular structure and behavior slightly. While for moistened flour, crystalline degree, gelatinization enthalpy, amylose leaching (AML) and falling number significantly decreased, but thermal transition temperatures increased with the rise of treating severity. Clumping of starch granules, aggregation of proteins and formation of amylose-lipid complexes occurred in both native and moistened flours. Broader pasting temperature ranges and higher viscosities were found on SS-modified flours. Additionally, SS treatment on moistened flours increased resistant and slowly digestible starch contents. In general, SS treatment induced changes in starch molecular structure and reactions among flour components leading to more stable structures, thus affecting their pasting behavior, thermal properties and in vitro digestion.     

Immunomodulatory activities of non-prolamin proteins in wheat germ and gluten

Albumin (Alb), globulin (Glo), glutelin (Gll) and glutenin (Gln) were separately extracted from wheat germ and wheat gluten. Amino acisd composition, molecular weight distribution, solubility, in vitro digestibility, and immunomodulatory activities were all analyzed. Gll and Gln have similar molecular weight distributions, which differed from those of Alb and Glo. Alb showed the highest solubility at various pH values (except pH 4.0), whereas Glo showed the highest in vitro digestibility. Glo and Gll have the highest proportion of essential to total amino acids, while Alb and Gll have the highest protein digestibility-corrected amino acid scores. Gll had the strongest immunomodulatory effects in terms of stimulation of RAW 264.7 cells to produce IL-6, TNF-α, and IL-10, and good stimulatory effects on splenocyte proliferation, production of IL-2, phagocytosis, and secretion of nitric oxide in RAW 264.7 cells. Gll can be considered a good protein source for use in health foods.     

Variability and cluster analysis of arabinoxylan content and its molecular profile in crossed wheat lines

Non-starch polysaccharides (NSP) in cereals, after starch and gluten proteins, determine the technological and nutritional properties of flour, dough and end product. From NSPs, the arabinoxylans are the most important components studied in wheat nowadays. The novelty of our study is the investigation of the variability and clustering ability of the wheat lines based on different quantitative and qualitative traits of arabinoxylans. The quantitative properties, the total and the water-extractable arabinoxylan content of wheat flours were measured separately by gas chromatography. The qualitative property referring to the structure of the molecule was the molecular weight distribution measured by size exclusion liquid chromatography. According to the results the variability of arabinoxylan properties in the breeding lines varied between wider ranges then the values measured for the parental varieties. The correlation between the quantitative parameters, and a trend between quantitative and qualitative parameters were described. During the cluster analysis, the parents were well separated into different groups. The parameters that played an important role in the clustering were the ratio of the probability of occurrence of molecular size ranges along with the quantitative traits of AX. The selected samples can be used for further targeted breeding while the methodology was used in this work can be suitable for selecting lines/varieties for special food or other industrial applications.     

Genetic responses in milling,flour quality,and wheat sensitivity traits to grain yield improvement in U.S. hard winter wheat

A rising global population necessitates continued genetic improvement of wheat (Triticum spp.), but not without monitoring of unintended consequences to processors and consumers. Our objectives were to re-establish trends of genetic progress in agronomic and milling traits using a generational meter stick as the timeline rather than cultivar release date, and to measure correlated responses in flour quality and human wheat-sensitivity indicators. Grain yield and kernel size showed stepwise increases over cycles, whereas wheat protein content decreased by 1.1 g/100 g. Reduced protein content, however, did not result in lower dough strength pertinent to bread baking. A novel method of directly testing gluten elasticity via the compression-recovery test indicated a general increase in gluten strength, whereas the ratio of total polymeric to total monomeric proteins remained stable. Also showing no change with genetic progress in yield were flour levels of gluten epitopes within the key immunotoxic 33-mer peptide. The oligosaccharide fructan, present in milled and wholemeal flours, increased with increasing grain yield potential. While yield improvement in U.S. bread wheat was not accompanied by a decline in gluten strength or systematic shift in a key wheat sensitivity parameter, the unanticipated rise in total fructans does implicate potentially new dietary concerns.     

Responses of glutamine synthetase activity and gene expression to nitrogen levels in winter wheat cultivars with different grain protein content

Glutamine synthetase (GS) plays a central role in plant nitrogen (N) metabolism, which improves crops grain protein content. A pot experiment in field condition was carried out to evaluate GS expression and activity, and grain protein content in high (Wanmai16) and low grain protein (Loumai24) wheat cultivars under two N levels (0.05 and 0.15 g N kg⁻¹ soil). High nitrogen (HN) resulted in significant increases in GS1 and GS2 expression at 10 days after anthesis (DAA), and higher GS activity during the entire grain filling stage. HN also significantly increased yield, grain protein content and protein fraction (except for glutenin of Luomai24) in two wheat cultivars, which indicated that it increased grain yield and protein content by improving nitrogen metabolism. Wanmai16 showed higher grain protein content, gliadin and glutenin content, and had higher expression level of GS2 both in flag leaves and grains at early grain filling stage. However, Luomai24 had greater yield and higher expression level of GS1. The difference expression of GS2 and GS1 genes indicates they had various contributions to the accumulation of protein and starch in wheat grains, respectively. The results suggest that GS2 would be serving as a potential breeding target for improving wheat quality.     

Effect of physicochemical and empirical rheological wheat flour properties on quality parameters of bread made from pre-fermented frozen dough

The objective of this study was to examine the influence of flour quality on the properties of bread made from pre-fermented frozen dough. The physicochemical parameters of 8 different wheat flours were determined, especially the protein quality was analysed in detail by a RP-HPLC procedure. A standardized baking experiment was performed with frozen storage periods from 1 to 168 days. Baked bread was characterised for specific loaf volume, crumb firmness and crumb elasticity. The results were compared to none frozen control breads. Duration of frozen storage significantly affected specific loaf volume and crumb firmness. The reduction of specific loaf volume was different among the used flours and its behaviour and intensity was highly influenced by flour properties. For control breads wet gluten, flourgraph E7 maximum resistance and RVA peak viscosity were positively correlated with specific loaf volume. However, after 1–28 days of frozen storage, wet gluten content was not significantly influencing specific loaf volume, while other parameters were still significantly correlated with the final bread properties. After 168 days of frozen storage all breads showed low volume and high crumb firmness, thus no significant correlations between flour properties and bread quality were found. Findings suggest that flours with strong gluten networks, which show high resistance to extension, are most suitable for frozen dough production. Furthermore, starch pasting characteristics were also affecting bread quality in pre-fermented frozen dough.     

Impact of lipases with different substrate specificity in wheat flour separation on the properties of the resultant gluten

Wheat gluten was isolated in a laboratory dough-batter flour separation process in the presence or absence of lipases differing in hydrolysis specificity. The obtained gluten was blended with wheat starch to obtain gluten-starch (GS) blends of which the water and oil binding capacities were investigated. Furthermore, GS blends were mixed into dough and processed into model breads, of which dough extensibility and loaf volume were measured, respectively. In comparison to GS blends prepared with control gluten, oil binding capacity was higher when GS blends contained gluten isolated with Lecitase Ultra (at 5.0 mg enzyme protein/kg flour), a lipase hydrolyzing both non-polar and polar lipids. Additionally, dough extensibility and total work needed for fracture were lower for dough prepared from GS blends containing gluten isolated with Lipolase (at 5.0 mg enzyme protein/kg flour), a lipase selectively degrading non-polar lipids. In GS blend bread making, this resulted in inferior loaf volumes. Comparable GS blend properties were measured when using control gluten and gluten isolated with YieldMAX, a lipase mainly degrading N-acyl phosphatidylethanolamine. In conclusion, properties of GS blend model systems are altered when gluten prepared in the presence of lipases is used to a degree which depends on lipase specificity and concentration.